Please note that 20µl of 5X Laemmli buffer were added per 100µl of tissue lysate, as a result the lysate may not be suitable for applications which require native protein in a non-denaturing buffer. The final Laemmli buffer concs are: 31.25mM Tris HCl, pH 6.8, 12.5% Glycerol, 1% SDS, 0.005% Bromophenol Blue, 25mM TCEP.

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Purification NotesTissue specimens are homogenized in modified RIPA buffer to obtain the soluble proteins, and centrifuged to clarify. The lysate solution may appear turbid at cold temperatures due to insolubility of buffer components. The solution should clear upon warming to room temperature.